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Storage and analysis of samples of water, fish, and mud from environments contaminated with abate.

Methods for extraction, cleanup, and analysis of samples of water, mud, and fish containing trace quantities of Abate have been developed. Water was extracted by high-speed stirring of 10 ml of hexane in a 300-ml sample. The extracts were evaporated and analyzed by gas chromatography with a limit of detection of 0.00003 ppm. Dried mud samples were extracted by shaking with acetone. An aliquot of the acetone extract was diluted with water and the Abate extracted into 10 ml of hexane by high-speed stirring. The extracts were analyzed by gas chromatography. Fish were extracted with methylene chloride, cleaned up on a silica gel column, and analyzed by gas chromatography. The limit of sensitivity of the methods for mud and fish was found to be 0.001 ppm. Fish samples were stored for 3 weeks in 10% formalin containing 5% sodium thiosulfate without significant loss of Abate residues. A biological magnification of greater than 100 was observed in fish exposed to Abate for 16 hr at concentrations of 0.02 and 0.002 ppm.

Animals↗

Sensitivity and accuracy of a bio-assay for the determination of the concentration of residual pesticide in natural water bodies.

Determination of pesticide levels in natural water bodies requires complex chemical methods not always available in tropical countries where such control programmes are implemented. Apart from chemical methods, effectiveness of treatment can be estimated by mortality of target and non-target species. But the latter criteria do not permit a good appraisal of dispersion and residual activity of pesticide. We propose here a simple and cheap bio-assay adaptable to water-borne vector or intermediate host control programmes. Results obtained with the bio-assay were compared with those obtained with gas liquid chromatography, a standard chemical method.

Aedes↗

The toxic effects of triorganotin(IV) compounds on the culicine mosquito, Aedes aegypti (L).

The structure-activity relationships of 25 triorganotin(IV) compounds were studied using the 4th instar A. aegypti larvae of the susceptible Liverpool Red-eye strain. The LC50 values of these compounds were compared with those of Malathion, DDT and Temephos. Maximal activities were obtained with tritolyltin chloride and its complexes with triphenyl-phosphine oxide and pyridine N-oxide, dimethyloctyltin- and diethyloctyltin acetates and tributyltin sucrose phthalate. A limited number of the triorganotin compounds was also tested with observed high activity against the 4th instar larvae of a local DDT-tolerant strain and against the 2nd instar larvae and adults of the susceptible strain. Delayed effect studies with both 2nd and 4th instar larvae showed high post-treatment mortality for the latter and, among the range of active compounds, the methyl- and aryltin compounds, in particular, were also found to exhibit high residual activities (100% kill) in laboratory tests conducted up to 10 weeks.

Aedes↗

A comparison of the effects of organophosphate insecticide exposure and temperature stress on fluctuating asymmetry and life history traits in Culex quinquefasciatus.

Effects of exposure to a non-chemical (temperature) or chemical (organophosphate insecticide) stressor during larval development were compared in Culex quinquefasciatus mosquitoes. Stress was measured in two ways: using conventional life history traits (survival, development time and a measure of body size) and by calculating the degree of developmental instability from the departure from bilateral symmetry of wing characters (fluctuating asymmetry). Increasing insecticide dose, but not temperature, was observed to elevate wing fluctuating asymmetry in male but not female mosquitoes. Insecticide treatment reduced survival and was associated with a significant reduction in wing trait sizes in both females and males but did not significantly affect development time. Temperature was associated with a significant reduction in all life history traits in both sexes. Therefore wing fluctuating asymmetry in C. quinquefasciatus cannot be used as a general biomonitor of all stress, although it may have potential as a more specific monitor of chemical stress. It needs to be complimented with other measures such as life history and biochemical methods. The significant differences in response between sexes may impact on results of short-term larval exposures to insecticides.

Animals↗

Esterases as biomarkers in Nereis (Hediste) diversicolor exposed to temephos and Bacillus thuringiensis var. israelensis used for mosquito control in coastal wetlands of Morbihan (Brittany, France).

Since 1998, a biomonitoring programme has been implemented to assess the potential impact of chemical mosquito control on macroinvertebrates of the coastal wetlands of Morbihan (Brittany, France). Acetylcholinesterase and carboxylesterases were used as biomarkers to assess the effects of Abate 500e (a.i. temephos) and Vectobac 12 AS (a.i. endotoxins of Bacillus thuringiensis var. israelensis, Bti) in Nereis (Hediste) diversicolor. Esterase inhibition revealed a marked impact of temephos, suggesting preferential contamination of the worms through the food. In Bti-exposed N. diversicolor, random variations of esterase activities were observed, that could not be attributed to the larvicide. However, esterases only reflected indirect physiological effects of Bti, and further investigations are needed to identify biomarkers more specific of Bti endotoxins.

Animals↗

Factors influencing the toxicity of xenobiotics against larval mosquitoes.

In order to examine the factors influencing xenobiotic toxicity against larval mosquitoes, the larvicidal performances of two conventional insecticides (temephos and Bacillus thuringiensis var. israelensis: Bti) and a new potential phyto-insecticide (decomposed leaf litter) were compared under different conditions against three detritivorous larval mosquito types. Bioassays performed under standard conditions indicated differential tolerance levels according to the xenobiotic and the larval type. Bioassays performed under different conditions of xenobiotic dose and geometry of the water column indicated differential effects of those parameters on mortality rates. This allowed us to distinguish the performances of temephos versus those of Bti and leaf litter. These toxicological performances were examined as indicators for analysis of xenobiotic bioavailability for mosquito larvae in environmental water, and also for their comparative interest in field mosquito control.

Aedes↗

The independent gene amplification of electrophoretically indistinguishable B esterases from the insecticide-resistant mosquito Culex quinquefasciatus.

Resistance to organophosphates in Culex mosquitoes is typically associated with increased activity of non-specific esterases. The commonest phenotype involves two elevated esterases, A2 and B2, while some strains have elevation of esterase B1 alone. Overexpression of the two B esterase electromorphs is due to gene amplification. Full-length cDNAs coding for amplified esterase B genes from a resistant Cuban strain (MRES, with amplified B1 esterase) and a Sri Lankan strain (PelRR, with amplified B2 esterase) of C. quinquefasciatus have been sequenced. In addition, a partial-length cDNA coding for a B esterase from an insecticide-susceptible Sri Lankan strain (PelSS) has been sequenced. All the nucleotide sequences and the inferred amino acid sequences show a high level of identify (> 95% at the nucleotide and amino acid level), confirming that they are an allelic series. The two B1 esterase nucleotide sequences (MRES and the previously published TEM-R [Mouches, Pauplin, Agarwal, Lemieux, Herzog, Abadon, Beyssat-Arnaouty, Hyrien, De Saint Vincent, Georghiou and Pasteur (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 2574-2578]) showed the lowest identity, and restriction-fragment-length-polymorphism analysis of the two strains was different. On the basis of these data we suggest that the two electrophoretically identical B1 esterase isoenzymes from California and Cuba have been amplified independently. Alternatively, if amplification has occurred only once, the original amplification has not occurred recently.

Amino Acid Sequence↗

Temephos-resistant larvae of Simulium sanctipauli associated with a distinctive new chromosome inversion in untreated rivers of south-western Ghana.

Larvae of the Simulium damnosum Theobald complex (Diptera: Simuliidae) were sampled in June 1996 from two sites in south-west Ghana where larviciding has not been applied: Sutri Rapids on the Tano river (05 degrees 23 minutes N 02 degrees 38 minures W) and Sekyere-Heman on the Pra river (05 degrees 11 minutes N 01 degrees 35 minutes W). All specimens were identified as Simulium sanctipauli Vajime & Dunbar sensu stricto (Diptera: Simuliidae). Bioassays with temephos (organophosphorus larvicide employed by the Onchocerciasis Programme for systematic treatment of most rivers across West Africa since the 1970s) showed about five-fold resistance in the Tano population (LC95 2.37-3.14 mg/L) and slight tolerance to temephos in the Pra population (LC95 0.67-0.76 mg/L), vs. the diagnostic concentration of 0.625 mg/L. Larval salivary polytene chromosomes of S. sanctipauli showed fixed inversions 1S-24/24, standard IIL-6 and a new inversion IL/36 polymorphism at Sutri on the Tano. These karyotype characteristics differ from those of temephos-resistant S. sanctipauli in rivers of C te d'Ivoire and other sites on the Tano in Ghana. Thus, temephos resistance in S. sanctipauli at Sutri is associated with distinct chromosomal configurations, showing that immigration was unlikely. This resistance could have been locally selected by exposure of S. sanctipauli larval populations to agrochemicals run-off from cocoa, coffee and oil plantations flanking the rivers.

Animals↗

A small scale field trial with Temephos (Abate) for the control of four species of Mansonia (Mansonioides) (Diptera: Culicidae) in Thailand.

A 60-week control experiment with Temephos (Abate) against a largely isolated population of Mansonia annulata, Ma. annulifera, Ma. indiana and Ma. uniformis was carried out in a pool of 1.2 ha area and average depth of over 2 m in southern Thailand. Floating paper tea bags containing 24 g of 1% Abate sand granules were applied in two rounds at a density of 1 bag 4 m-2. Each round consisted of five applications at intervals of 13 days. During the application rounds mosquito larvae disappeared completely from the pool. This apparent effect of Abate was confirmed by bioassays made with pool water and larvae of Culex quinquefasciatus. Inactivation of Abate was linear during one week, with a slope of 5.8% per day. Adult densities dropped during the application rounds and, over a period of 48 weeks, were 78-85% less than during the corresponding time of the preceding reference year. The annual pattern of adult abundance that normally shows distinct seasonal peaks was strongly altered, although meterological data were comparable to the reference year.

Animals↗

Successful control of onchocerciasis vectors in San Vicente Pacaya, Guatemala, 1984-1989.

Between 1984 and 1989, the onchocerciasis-vector control zone on the pilot area of San Vicente Pacaya, Guatemala, which had been subject to experimental control since 1979, was extended from 91.3 to 148.6 km2. Temephos was used as a larvicide against Simulium ochraceum s.l. the target species. As a new strategy, only breeding sites with water discharges of 0.1-10 litres/s were treated, every 2 weeks. This approach provided a substantial reduction in effort, number of treated sites, time and cost. To assess the effect of the temephos, nine sites were selected in which standardized collections of adult Simulium were made twice a month, by human bait. There was an obvious difference between the pre- and post-control mean densities of flies at each site. In the northern area, which includes the Lavaderos, Barretal, Colina and Rodeo sites, the biting density in 1979, before treatment, varied between 10 and 64 flies/man-hour (FMH). Four years later, this had been reduced to 0.1-3.2 FMH, and by the end of the present study, in 1989, the mean density was zero FMH. In the southern area, which lies south-east of Lavaderos (and includes Guachipilin, Ingerto, Pena Blanca and Sierra Morena), the density during the pre-control phase was 24 FMH at one of the two sites investigated at the time and 39.3 FMH at the other. It fell to 0.1-0.5 FMH after 5 years of control and to zero (three sites) or close to zero (< or = 0.5 FMH; one site) for the last 4 years of the present study. To assess the effect of vector control on onchocerciasis prevalence and incidence, 1280 residents from six endemic communities, out of 12,000 permanent inhabitants, were examined. In Santa Cruz, Patrocinio and Los Rios, the prevalence of skin microfilariae in the subjects from each community fell from 8.1%-37.8% during the pretreatment, base-line period to 0.0%-31.5% when the study foci were totally integrated into the vector-control operation following treatment. Incidence among children (aged < or = 9 years) varied from 0%-25% for the period 1982-1984 but, thereafter, not a single case appeared in four of the six study communities (Santa Cruz, Patrocinio, Los Rios and Berlin). Incidence in Guachipilin did not decline appreciably, probably because of human migration into the area from other onchocerciasis foci. The prevalence of nodules followed a similar trend to those of the prevalence and incidence of skin microfilariae, falling from 9.1%-45.0% pre-control to 1.8%-14.3% 10 years later.

Animals↗

[The initial activity and persistance of an experimental larvicide with a base of Bacillus thuringiensis var. israelensis and a commercial preparation containing the organophosphorus insecticide temephos].

The larvicide activity of the experimental preparation SAN 402 I WDC containing B. Thuringiensis var. Israelensis and that of Abate 500 E (44 % temephos) were tested on the larvae of Aedes aegypti. The studies were performed on the following media: distilled water, pure and buffered at pH 9, 7 and 4 and pond water whether or not free from materials in suspension. The activity of both preparations is not influenced by the composition of the media, exception made for the presence of materials in suspension. The persistance varies according to the kind of the media and the concentration level. It is in general rather low. Both preparations behave rather similar, the microbial insecticide however is more influenced by the presence of materials in suspension.

Aedes↗

Eliminating onchocerciasis after 14 years of vector control: a proved strategy.

From 1976 through 1989, weekly aerial spraying operations against blackflies were carried out along the rivers of a wide savanna area of West Africa (approximately 700,000 km(2)) where onchocerciasis was hyperendemic. The level of endemicity began to decrease significantly after 4 years of vector control and became very low in 1989. This situation has been maintained without any vector control activity or chemotherapy, and no incidence of any new cases has been detected. An ophthalmological study carried out in 2000 has confirmed these good results, showing only cicatricial ocular lesions in the examined population. These results led to the conclusion that 14 years of vector control may achieve long-term elimination of onchocerciasis, even in the absence of chemotherapy, provided that the treated areas are not subjected to any contamination by exogenous parasites carried in infected humans or flies.

Adult↗